Este trabajo fue apoyado por el suizo nacional ciencia Fundación beca 150271.

este protocolo experimental ha sido aprobado por la Oficina veterinaria Cantonal, Fribourg y el suizo Federal veterinario oficina, Suiza (FR 2015/58).
 Nota: se utilizaron conejos macho NZB entre 2.8 a 3.2 kg. Los animales fueron alojados bajo condiciones convencionales (12 h de luz y oscuro ciclo, proporcionada alimento y agua ad libitum). Antes de denudación del globo, los animales se aclimataron durante 1 semana durante la cual fueron alimentados con dieta normal chow. Después de una semana de aclimatación, conejos fueron cambiados a aterogénico dieta consistente en grasas (8.6%) y saturan ácidos grasos con 205 mg/kg dieta de colesterol (1%) para la duración del estudio entero. Lesión de globo en la arteria ilíaca izquierda se realizó 1 semana después de la iniciación de la dieta y los animales fueron sacrificados después de 2 semanas o 4 semanas de lesión de globo.
 1. procedimientos preoperatorios <ol><li>esterilizar todos los instrumentos quirúrgicos antes de usar con un esterilizador de bolas de vidrio u otro instrumento adecuado.</li>
	<li>Preparar y comprobar el conjunto de catéter balón.
	<ol><li>Coloque una cerradura de luer 1 mL jeringa llena con solución salina normal a la parte de la cerradura del luer del catéter con balón. Vigilar cuidadosamente por ausencia de aire atrapado. Compruebe las fugas y garantizar la inflación adecuada globo presionando el émbolo de la jeringa de.</li>
	</ol></li> <li>Pesan el conejo y encienda el thermopad a 37 ° C.</li>
	<li>Utilizar una solución de buprenorfina a una concentración de 0,3 mg / ml. inyectar por vía subcutánea una dosis de 0,01 mg/kg.</li>
	<li>Anesthetize el conejo con 5% isoflurano y 5 L/min O 2 en una cámara de inducción de 10-15 min.</li>
	<li>Poner el conejo anestesiado en la almohada en la plataforma quirúrgica. Coloque el parche y clips para monitorear la temperatura, respiración y electrocardiograma.</li>
	<li>Colocar el hocico del conejo a una mascarilla conectada a una máquina de anestesia adecuada. Mantener la anestesia con isoflurano (4.0% con 2,5 L/min O 2). Confirmar la anestesia apropiada (indicada por la falta de tono muscular y pérdida de los reflejos nauseoso y pinnae).</li>
	<li>Aplique pomada oftálmica en ambos ojos para evitar que la córnea se seque. Cuelgue el conejo con una hoja quirúrgica estéril con sólo el miembro inferior expuesto.</li>
</ol> 2. Protocolo quirúrgico <ol><li>eliminar el pelo de la zona ventral justo debajo de las articulaciones de rodilla con cortapelos animales.</li>
	<li>Limpiar el área con desinfectante adecuado para limpiar la piel y quitar el pelo flojo.</li>
	<li>Localizar la arteria safena y haga una incisión de piel pequeña de unos 1,5 cm de longitud con un bisturí.</li>
	<li>Exponer una pequeña porción de la arteria safena con pinzas curvas pequeñas, sin dañar la vena femoral y nervio femoral.</li>
	<li>Coloque dos lazos de ligadura suelta (5-0 seda) debajo de la arteria safena y ATA una ligadura hacia el extremo distal de la arteria. Colocar un clamp microvascular por encima de la ligadura para detener el flujo de sangre desde la arteria ilíaca.</li>
	<li>Por vía tópica se aplica una gota de la papaverina para dilatar la arteria y evitar vasoespasmo.</li>
	<li>Levante la arteria safena con la ayuda de la ligadura atada y hacer una incisión pequeña arteriotomía utilizando 24 aguja g.</li>
	<li>Elevar las aletas de la incisión con unas pinzas finas y lentamente introducir una selección de vena o una aguja guía en el lumen de la arteria.</li>
	<li>Insertar un catéter de embolectomía arterial de Fogarty francés 2 dentro de la arteria safena. Quite la selección de la vena y pinzas microvasculares.</li>
	<li>Haga avanzar el catéter hasta la sexta marca (20-25 cm) correspondiente a una posición aproximadamente 2-5 cm por encima de la bifurcación ilíaca.</li>
	<li>Inflar el balón con solución salina normal 0,1 mL usando una jeringa de 1 mL o a una presión nominal de 6 atm con un inflador manual regulada tal como se describe en el 16 , 22.</li>
	<li>Mantenga el catéter de balón con pinzas y arrastre por 6 cm a través de la arterytoward ilíaca el punto de inserción, mientras gira el catéter.</li>
	<li>Desinflar el balón tirando hacia atrás el émbolo de la jeringa de.</li>
	<li>Repita 2.10 a 2.13 tres veces para asegurar la completa denudación endotelial.</li>
	<li>Retirar el catéter e inmediatamente atar el lazo de la ligadura por encima del sitio de arteriotomía para detener el sangrado.</li>
	<li>Aplique todo antiséptico adecuado alrededor de la periferia de la herida y limpiar lejos los coágulos de sangre. Cerrar la incisión de la piel con una sutura 5-0 y desinfectar el sitio de la cirugía con solución de povidona-yodo.</li>
	<li>Repita 2.1 a 2.16 en el contralateral ilíaca usando un catéter nuevo.</li>
	<li>Esponja el ungüento oftálmico de ojos.</li>
</ol> 3. Cuidados postoperatorios <ol><li>administrar sulfadoxina 40 mg/kg y el trimethoprim 8 mg/kg o cualquier otro antibiótico adecuado inmediatamente después del procedimiento quirúrgico.</li>
	<li>Durante el período de recuperación anestésica, mantener el conejo sobre una almohadilla de calor colocada en una jaula limpia esterilizada.</li>
	<li>Quitar el parche monitoreo y clips de.</li>
	<li>Después de la recuperación, regresar los conejos en sus jaulas hogar. Inyectar en forma subcutánea buprenorfina 0.05 - 0.1 mg/kg post - operativamente cada 6-12 h para 48 h. continuar dieta aterogénico por otro dos semanas o cuatro semanas.</li>
</ol> 4. La cosecha de tejido y análisis de composición de la placa <ol><li>después de dos semanas (para la placa delgada temprano) o de tres semanas de lesión de globo, anestesiar al conejo con isoflurano en una manera similar como se describe arriba.</li>
	<li>Abrir la cavidad torácica y eutanasia a los conejos por intracardial exsanguination.</li>
	<li>Aislar las arterias ilíacas, como se describe en el 23.
	<ol><li>Brevemente, abrir el abdomen y exponer el retroperitoneum. Seguimiento de la aorta hacia la bifurcación ilíaca y atarla por encima de la bifurcación. Retire con cuidado los tejidos circundantes para denunciar y aislar ambas arterias ilíacas.</li>
	</ol></li> <li>Diseque hacia fuera ambas arterias ilíacas y sumergirlos en solución salina amortiguada de fosfatos helada. Eliminar los coágulos con la ayuda de fórceps. Cada arteria ilíaca se dividen en segmentos de 4 a 6 para caracterizar el espesor de la placa a lo largo de la arteria.</li>
	<li>Inmediatamente incrustar los segmentos arteriales en un molde con temperatura óptima de corte compuesto, snap-congelación con nitrógeno líquido y guardar a-70 ° C. preparar 5 μm secciones de espesor con un criostato como se describe en 24.</li>
	<li>Realizar histología, la inmunofluorescencia o la coloración immunohistochemical para morfometría, placa lípidos y contenido celular como se describe en 10 , 25. 
	Nota: Brevemente, enjuagar las secciones arteriales con tampón fosfato salino (PBS) y permeabilizar utilizando 0,2% Tritón. Enjuague las secciones con PBS y bloquear sitios no específicos con seroalbúmina bovina al 2% durante 30 minutos incubar las secciones por 1 h a 37 ° C con anti actina α-SM (1: 200) o RAM11 anticuerpo (1: 200). Enjuague las secciones con PBS e incubar con el anticuerpo secundario apropiado durante 30 min a 37 ° C. lavado nuevamente con PBS y añadir Hoechst (5 μg/mL) durante 10 min detectar núcleos.</li>
</ol>

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J., Bao, S., Celermajer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography+for+serial+in+vivo+imaging+of+aortic+plaque+in+the+rabbit:+a+preliminary+experience.">Optical coherence tomography for serial in vivo imaging of aortic plaque in the rabbit: a preliminary experience.</a> Open Heart. 2, e000314 (2015).</li><li>Abela, O. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plaque+Rupture+and+Thrombosis:+the+Value+of+the+Atherosclerotic+Rabbit+Model+in+Defining+the+Mechanism.">Plaque Rupture and Thrombosis: the Value of the Atherosclerotic Rabbit Model in Defining the Mechanism.</a> Curr Atheroscler Rep. 18, 29 (2016).</li><li>Yamashita, A., Asada, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rabbit+model+of+thrombosis+on+atherosclerotic+lesions.">A rabbit model of thrombosis on atherosclerotic lesions.</a> J Biomed Biotechnol. 2011, 424929 (2011).</li></ol>

Los autores no declaran a intereses financieros en competencia.

El modelo de aterosclerosis de la arteria ilíaca de conejo es ampliamente utilizado en la investigación de la aterosclerosis. Con este protocolo los conejos desarrollaron rápidamente las placas más severas y avanzadas en comparación con las lesiones espontáneas convertidas sólo dieta de colesterol. Lo importante, animales recuperan rápidamente de la cirugía.
El principal estímulo de la aterogénesis es el daño mecánico causado por el catéter de balón que lesiona el endotelio y el...

La ruptura de las placas ateroscleróticas vulnerables es una de las principales causas de muerte en los países industrializados1. Aunque la investigación en las últimas décadas ha desarrollado varios mecanismos moleculares y celulares implicados en la progresión de la placa, continuó todavía se necesitan esfuerzos no sólo para desentrañar el complejo mecanismo de progresión de la enfermedad sino también para probar nuevas terapéuticas se acerca. Se han propuesto varios modelos animales para el estudio de la aterosclerosis. Manipulación genética, lesión de endotelio de alimentación o mecánico de colesterol son las estrategias estándar compartidas por modelos más animales de la aterosclerosis, incluyendo ratones, conejos o minipigs. Entre éstos, NZB los conejos son sensibles a la dieta de colesterol mientras que los ratones y las ratas normales no absorben mucho colesterol de la dieta2,3,4. Conejos desarrollan espontáneamente las lesiones aórticas ricas en macrófagos con algún componente fibroso cuando alimentados con colesterol dieta rica5,6. Sin embargo, el tiempo largo preparatorio de 4 a 8 meses para inducir la aterosclerótica plaquesby alimentación colesterol dieta solo6,7 es un gran inconveniente para la mayoría de los ajustes experimentales. En búsqueda para inducir lesiones en relativamente poco tiempo, una combinación de colesterol alto dieta y balón la lesión ha sido desarrollada por Baumgarter y Studer8. El objetivo general de esta técnica es inducir a las placas ateroscleróticas compuestas de células espumosas (similares a la estría grasa en los seres humanos) en conejos hipercolesterolémicos dentro de 2 semanas. La presente técnica describe el procedimiento de lesión de la pared arterial basado en método de Baumgarter utilizando un catéter con balón en la arteria ilíaca de conejos hipercolesterolémicos NZB.
Junto con una dieta rica en colesterol, lesiones resultantes de globo inducido la endotelialización conducirá a la aterosclerosis. Lesión de globo acelera la formación de lesiones ateroscleróticas y produce placa de tamaño uniforme y distribución. Engrosamiento intimal aumenta durante un período de tiempo y célula intimal infiltración comienza en pocos días después de lesión. Grasos con macrófagos importantes comienzan a aparecer después de 7-10 días de lesión de globo y están representados como lesión tipo II según la clasificación de la Asociación Americana del corazón. Lesión de globo en conejo se realiza con frecuencia en la aorta para estudiar la composición de la placa. El endotelio neointimal expresa altos niveles de la molécula de adhesión intercelular. Las placas se asocian a disección medial y cambios adventicial. Las lesiones ateroscleróticas están compuestos por lípidos, proliferación las células musculares lisas (SMCs), fibras de colágeno y células inflamatorias que se acumulan en el endotelio regenerado y son principalmente de tipo II en la naturaleza. La distribución topológica de las placas de conejo fue similar al reportado en aortas humanas 9,10 , en principio, la aorta es más grande en tamaño en comparación con las arterias ilíacas y produciría la placa de mayor longitud. Sin embargo, la gran ventaja de la utilización de la arteria ilíaca como el sitio de la aterosclerosis en conejos es su accesibilidad, su similitud en el contenido muscular de la arteria coronaria humana11, lesión uniforme desarrollo12, factor tisular alta actividad13 y recipiente consistente dimensión comparable a la coronaria humana permitiendo la evaluación de dispositivos comercialmente manufacturados morfométricas y criterios de valoración angiográficas. Se han investigado métodos invasivos y no invasivos para analizar las placas en las arterias ilíacas de conejo en el animal vivo. Informes anteriores describen el uso de resonancia magnética (MRI) con la ayuda de un Señor de 2,35-tesla sistema 14 además, el ultrasonido intravascular (IVUS) o catéteres de tomografía de coherencia óptica pueden ser adecuadamente aplicada a la imagen placas ateroscleróticas en las arterias ilíacas conejo. La arteria ilíaca es accesible para la proyección de imagen de ultrasonido cuando se usa una ecografía de alta resolución y la aorta puede estudiarse también con esta técnica.
En la última década, este modelo de conejo de lesión de globo ha ayudado a comprender aún más los mecanismos de progresión de placa15y regresión de placa16. Además, el modelo se ha utilizado para estudiar la influencia de nuevos agentes terapéuticos tales como las estatinas, antiagregantes estándar, antioxidante agentes17,18 y stents liberadores de fármacos como everolimus o liberador de zotarolimus stent19,20 en engrosamiento neointimal. Este modelo también se ha utilizado para investigar la proyección de imagen intravascular del catéter21la proyección de imagen de la fluorescencia del infrarrojo cercano.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>New Zealand White rabbits</td>
			<td>Charles River laboratories,France</td>
			<td>Cre:KBL(NZW)</td>
			<td></td>
		</tr>
		<tr>
			<td>Cholesterol rich diet</td>
			<td>Ssniff spezialdi&#228;ten</td>
			<td>Ssniff EF K High Fat and Cholesterol</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass bead sterilizer-Germinator 500</td>
			<td>VWR, Leicestershire, UK</td>
			<td>101326-488</td>
			<td></td>
		</tr>
		<tr>
			<td>Fogarty balloon embolectomy catheters, 2 French</td>
			<td>Edwards Lifesciences, Switzerland</td>
			<td>120602F</td>
			<td>For single use only</td>
		</tr>
		<tr>
			<td>Luer Lock Syringe</td>
			<td>Becton, Dickinson and Company, USA</td>
			<td>309628</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermopad Type 226</td>
			<td>Solis, Switzerland AG</td>
			<td>397387</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine- Temgesic</td>
			<td>Reckitt Benckiser AG, Switzerland</td>
			<td>7.68042E+12</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Piramal Critical Care, Inc, Bethlehem, PA 18017</td>
			<td>2667-46-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Anaesthesia machine-combi-vet Base Anesthesia System</td>
			<td>Rothacher Medical GmbH, Switzerland</td>
			<td>CV 30-301-A</td>
			<td></td>
		</tr>
		<tr>
			<td>Cardell touch veterinary vital signs monitor</td>
			<td>Midmark, Ohio, USA</td>
			<td>8013-001</td>
			<td></td>
		</tr>
		<tr>
			<td>Ophthalmic ointment-Humigel</td>
			<td>Virbac, France</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Animal hair clippers</td>
			<td>Aesculap AG, Germany</td>
			<td>GT420</td>
			<td></td>
		</tr>
		<tr>
			<td>Disinfectant-Betadine solution</td>
			<td>MundipharmaMedicalCompany, Switzerland</td>
			<td>14671-1203</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #7 Forceps</td>
			<td>FST Germany</td>
			<td>11274-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Medium and small microscissors</td>
			<td>Medline International Switzerland S&#224;rl</td>
			<td>UC4337</td>
			<td></td>
		</tr>
		<tr>
			<td>Microvascular clamps</td>
			<td>FST, Germany</td>
			<td>18051-28</td>
			<td></td>
		</tr>
		<tr>
			<td>Papaverine</td>
			<td>ESCA chemicals, Switzerland</td>
			<td>RE 356 803</td>
			<td></td>
		</tr>
		<tr>
			<td>Vein Pick</td>
			<td>Harvard Apparatus, Cambridge, UK</td>
			<td>72-4169</td>
			<td>For single use only</td>
		</tr>
		<tr>
			<td>Saline</td>
			<td>Laboratorium Dr. G. Bichsel AG, , Switzerland</td>
			<td>1330055</td>
			<td></td>
		</tr>
		<tr>
			<td>Polysorb 5-0 suture</td>
			<td>Covidien AG, Switzerland</td>
			<td>UL 202</td>
			<td>Monofilament</td>
		</tr>
		<tr>
			<td>Sulfadoxine and Trimethoprim-Trimethazol</td>
			<td>Werner Stricker AG, Switzerland</td>
			<td>Swissmedic Nr. 50&#39;361</td>
			<td></td>
		</tr>
		<tr>
			<td>Antiseptic- Octenisept</td>
			<td>Sch&#252;lke &#38; Mayr AG, Switzerland</td>
			<td>GTIN: 4032651214068</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline</td>
			<td>Roth</td>
			<td>1058.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Isobutanol-2-Methylbutane</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>M32631-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>Optimum Cutting Temperature compound-Tissue-Tek</td>
			<td>VWR Chemicals, Belgium</td>
			<td>25608-930</td>
			<td></td>
		</tr>
		<tr>
			<td>Cryostat</td>
			<td>Leica, Glattbrugg, Switzerland</td>
			<td>Leica CM1860 UV</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass slide- Superfrost Plus</td>
			<td>Thermo Scientific</td>
			<td>4951PLUS4</td>
			<td></td>
		</tr>
		<tr>
			<td>Mayer&#39;s Haematoxylin</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>MHS32-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>Eosin 0.5% aq.</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>HT110232-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>Oil Red O</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>O0625-25G</td>
			<td></td>
		</tr>
		<tr>
			<td>&#945;-smooth muscle actin antibody</td>
			<td>Abcam, UK.</td>
			<td>ab7817</td>
			<td></td>
		</tr>
		<tr>
			<td>Macrophage Clone RAM11 antibody</td>
			<td>DAKO, Switzerland</td>
			<td>M063301</td>
			<td></td>
		</tr>
		<tr>
			<td>Hoechst</td>
			<td>Abcam, UK.</td>
			<td>ab145596</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat polyclonal Secondary Antibody (Chromeo 546)</td>
			<td>Abcam, UK.</td>
			<td>ab60316</td>
			<td></td>
		</tr>
		<tr>
			<td>Alexa Fluor 488/547</td>
			<td>Abcam, UK.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glycergel Mounting Medium, Aqueous</td>
			<td>DAKO, Switzerland</td>
			<td>C056330</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>H3136-25G</td>
			<td></td>
		</tr>
		<tr>
			<td>Ferric chloride for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>157740-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Iodine for Movat staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>207772-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium iodide for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>60400-100G-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcian blue for Movat staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>A5268-10G</td>
			<td></td>
		</tr>
		<tr>
			<td>Strong Ammonia for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>320145-500ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Brilliant crocein MOO for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>210757-50G</td>
			<td></td>
		</tr>
		<tr>
			<td>Acid Fuchsin for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>F8129-50G</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Thiosulfate for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>72049-250G,</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphotungstic acid for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>79690-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Crocin for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>17304-5G</td>
			<td></td>
		</tr>
		<tr>
			<td>EUKITT for Movat pentachrome staining</td>
			<td>Sigma-Aldrich, Switzerland</td>
			<td>03989-100ML</td>
			<td></td>
		</tr>
	</tbody>
</table>

the rabbit model of accelerated atherosclerosis: a methodological perspective of the iliac artery balloon injury

Síndrome coronario agudo resultante de la obstrucción coronaria después de la ruptura y desarrollo de la placa aterosclerótica es la causa principal de muerte en el mundo industrializado. Conejos Nueva Zelanda blanco (NZB) son ampliamente utilizados como modelo animal para el estudio de la aterosclerosis. Desarrollan lesiones espontáneas cuando alimentados con la dieta aterogénico; sin embargo, esto requiere mucho tiempo de 4 a 8 meses. Para mejorar y acelerar la aterogénesis, se emplea a menudo una combinación de dieta aterogénico y lesión endotelial mecánica. El procedimiento presentado en la inducción de las placas ateroscleróticas en conejos utiliza un catéter con balón para alterar el endotelio en la arteria ilíaca izquierda de NZB conejos alimentados con la dieta aterogénico. Daño mecánico causado por el catéter con balón induce una cadena de reacciones inflamatorias iniciar neointimal acumulación del lípido en una manera dependiente del tiempo. Placa aterosclerótica después de espesamiento de neointimal de mostrar globo lesiones con infiltración extensa de lípidos, contenido de la celda alta del músculo liso y presencia de macrófagos derivados de células de la espuma. Esta técnica es simple, reproducible y produce la placa de longitud controlada dentro de la arteria ilíaca. Todo el procedimiento se completa dentro de 20-30 minutos. El procedimiento es seguro con baja mortalidad y ofrece también gran éxito en la obtención de importantes lesiones intimales. El procedimiento del catéter de balón indujo resultados de lesión arterial en la aterosclerosis dentro de dos semanas. Este modelo puede utilizarse para investigar la patología de la enfermedad, diagnóstico por imágenes y evaluar nuevas estrategias terapéuticas.

Lesiones del globo de la arteria ilíaca fue realizada con éxito sin complicaciones (figura 1). El tiempo quirúrgico total varió de 20 a 30 min de lesiones realizado en sólo una arteria ilíaca y 35 a 45 min para las lesiones en ambas arterias. El conejo se recuperó dentro de 1 h después de lesión de globo. Todos los animales aparecieron sanos sin pérdida significativa de peso. Se ha encontrado ninguna infección, edema o trombosis arterial. El área ...

Watch this Scientific Journal Video about The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury at JoVE.com

The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

Modelos animales de aterosclerosis son esenciales para comprender el mecanismo e investigar nuevos enfoques para prevenir el desarrollo de la placa o ruptura, una causa principal de muerte en el mundo industrializado. Este protocolo utiliza una combinación de lesión de globo y dieta rica en colesterol para inducir a las placas ateroscleróticas en arteria ilíaca de conejo.

El modelo de conejo de ateroesclerosis acelerada: una perspectiva metodológica de la lesión de globo de la arteria ilíaca

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the ...

the-rabbit-model-accelerated-atherosclerosis-methodological

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

12.0K vistas.  University of Fribourg. Modelos animales de aterosclerosis son esenciales para comprender el mecanismo e investigar nuevos enfoques para prevenir el desarrollo de la placa o ruptura, una causa principal de muerte en el mundo industrializado. Este protocolo utiliza una combinación de lesión de globo y dieta rica en colesterol para inducir a las placas ateroscleróticas en arteria ilíaca de conejo.

Video: The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Un modelo murino de la implantación de un stent en la arteria carótida para el Estudio de la reestenosis

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Investigación

Medicina

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

Un modelo murino de isquemia-reperfusión miocárdica lesión a través de la ligadura de la arteria descendente anterior

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and cellular mechanisms involved in vascular smooth muscle dedifferentiation, neointima formation and vascular remodeling. Male Sprague-Dawley rats are the most frequently employed animals for this model. Female rats are not preferred as female hormones are protective against vascular diseases and thus introduce a variation into this procedure. The left carotid is typically injured with the right carotid serving as a negative control. Left carotid injury is caused by the inflated balloon that denudes the endothelium and distends the vessel wall. Following injury, potential therapeutic strategies such as the use of pharmacological compounds and either gene or shRNA transfer can be evaluated. Typically for gene or shRNA transfer, the injured section of the vessel lumen is locally transduced for 30 min with viral particles encoding either a protein or shRNA for delivery and expression in the injured vessel wall. Neointimal thickening representing proliferative vascular smooth muscle cells usually peaks at 2 weeks after injury. Vessels are mostly harvested at this time point for cellular and molecular analysis of cell signaling pathways as well as gene and protein expression. Vessels can also be harvested at earlier time points to determine the onset of expression and/or activation of a specific protein or pathway, depending on the experimental aims intended. Vessels can be characterized and evaluated using histological staining, immunohistochemistry, protein/mRNA assays, and activity assays. The intact right carotid artery from the same animal is an ideal internal control. Injury-induced changes in molecular and cellular parameters can be evaluated by comparing the injured artery to the internal right control artery. Likewise, therapeutic modalities can be evaluated by comparing the injured and treated artery to the control injured only artery.

This protocol uses a balloon catheter to cause an intraluminal injury on the rat carotid artery and henceforth elicit neointimal hyperplasia. This is a well-established model for studying the mechanisms of vascular remodeling in response to injury. It is also widely used to determine the validity of potential therapeutic approaches.

Lesión Vascular Globo y Administración intraluminal en rata arteria carótida

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and ...

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

Una rata carótida lesión con el balón modelo para probar Terapéutica Remodelación Anti-vasculares

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

Inducción de la Aterosclerosis Acelerada en Ratones: El Modelo de "Lesiones por Alambre"

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen. A short incubation period of the vector in the lumen, with subsequent aspiration of the lumen contents, is sufficient to achieve efficient and durable expression of the transgene in the endothelium, with no detectable transduction or expression outside of the isolated arterial segment. The method allows assessment of the biological activities of transgene products both in normal arteries and in models of human vascular disease, while avoiding systemic effects that could be caused either by targeting gene delivery to other sites (e.g. the liver) or by the alternative approach of delivering genetic constructs to the endothelium by germ line transgenesis. Application of the method is limited by the need for a skilled surgeon and anesthetist, a well-equipped operating room, the costs of purchasing and housing rabbits, and the need for expertise in gene-transfer vector construction and use. Results obtained with this method include: transgene-related alterations in arterial structure, cellularity, extracellular matrix, or vasomotor function; increases or reductions in arterial inflammation; alterations in vascular cell apoptosis; and progression, retardation, or regression of diseases such as intimal hyperplasia or atherosclerosis. The method also allows measurement of the ability of native and synthetic DNA regulatory sequences to alter transgene expression in endothelial cells, providing results that include: levels of transgene mRNA, levels of transgene protein, and levels of transgene enzymatic activity.

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

This method is to introduce a transgene into the endothelium of rabbit carotid arteries. Introduction of the transgene allows the assessment of the biological role of the transgene product either in normal arteries or disease models. The method is also useful for measuring activity of DNA regulatory sequences.

En Vivo Transferencia génica en el endotelio de la arteria carótida común conejo

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

Un modelo de conejo de expresión del transgen Durable en la vena yugular a los injertos de interposición de la arteria carótida común

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Medición de la contractilidad isométrica de la arteria mesentérica de ratón con cable miografía

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

Una lesión de globo segmentario controlada por presión arterial carótida de rata con aplicación terapéutica periavential

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

The Intra-Aortic Balloon Pump

Cardiogenic shock remains one of the most challenging clinical syndromes in modern medicine. Mechanical support is being increasingly used in the management of cardiogenic shock. Intra-aortic balloon pump (IABP) is one of the earliest and most widely used types of mechanical circulatory support. The device acts by external counterpulsation and uses systolic unloading and diastolic augmentation of aortic pressure to improve hemodynamics. Although IABP provides less hemodynamic support when compared with newer mechanical circulatory support devices, it can still be the mechanical support device of choice in appropriate situations because of its relative simplicity of insertion and removal, need for smaller size vascular access and better safety profile. In this review, we discuss the equipment, procedural and technical aspects, hemodynamic effects, indications, evidence, current status and recent advances in the use of IABP in cardiogenic shock.

We describe the steps for the percutaneous implantation of the intra-aortic balloon pump (IABP), a mechanical circulatory support device. It acts by counterpulsation, inflating at the onset of diastole, augmenting diastolic aortic pressure and improving coronary blood flow and systemic perfusion, and deflating before systole, reducing left ventricular afterload.